Fig. 5: Spindlin1-HBx promotes HBV gene transcription in HepG2-NTCP cells. | Nature Communications

Fig. 5: Spindlin1-HBx promotes HBV gene transcription in HepG2-NTCP cells.

From: Molecular insights into Spindlin1-HBx interplay and its impact on HBV transcription from cccDNA minichromosome

Fig. 5

a ChIP experiments using HBV infected cells showed Spindlin1 is recruited to the HBV cccDNA. b–e At 3 days post infection (dpi) with HBV, cells were transfected with control siRNA (Ctrl siRNA) or Spindlin1 siRNAs and collected at 10dpi for HBV RNA (b, c), HBeAg (d), and HBcAg (red, e) levels analysis. e Cell nuclei were stained with DAPI dye (blue). Scale bars: 100 μm. Three biological repeats of each experiment were repeated independently with similar results. f–g Ectopic expression of wild-type Spindlin1 (SPIN1) restored the HBV transcription (f) and HBeAg (g) levels in Spindlin1 knockdown cells. h HepG2-NTCP cells were infected with normalized amount of wild-type HBV (HBV WT) or HBx-deficient HBV (HBV X-). At 7dpi, cells were harvested and analyzed by ChIP using antibodies against Spindlin1. i–j Ectopic expression of wild-type Spindlin1 but not its H3K9me3 or H3K4me3 or double-binding deficient mutant restored HBV expression (i) and HBeAg (j) levels in Spindlin1 knockdown cells. EV, empty vector. Data represent the mean ± SD (n = 3 independent experiments). P-values between the groups were calculated with an unpaired two-tailed t test. Source data are provided as a Source Data file.

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